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用 TRIzol 分离得到的蛋白质与二维电泳和质谱分析兼容。

Proteins isolated with TRIzol are compatible with two-dimensional electrophoresis and mass spectrometry analyses.

机构信息

Institute of Environmental Science and Research, Kenepuru Science Centre, Porirua, New Zealand.

出版信息

Anal Biochem. 2012 Feb 1;421(1):330-2. doi: 10.1016/j.ab.2011.10.045. Epub 2011 Nov 4.

Abstract

TRIzol is used for RNA isolation but also permits protein recovery. We investigated whether proteins prepared with TRIzol were suitable for two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry. Proteins from TRIzol-treated SH-SY5Y cells produced 2-DE spot patterns similar to those from an equivalent untreated sample. Subsequent identification of TRIzol-treated proteins using peptide mass fingerprinting was successful. TRIzol exposure altered neither the mass of myoglobin extracted from sodium dodecyl sulfate (SDS) gels nor the masses of myoglobin peptides produced by in-gel trypsin digestion. These findings suggest that proteins isolated with TRIzol remain amenable to proteomic analyses.

摘要

TRIzol 用于 RNA 分离,但也允许回收蛋白质。我们研究了用 TRIzol 制备的蛋白质是否适合二维凝胶电泳 (2-DE) 和基质辅助激光解吸/电离质谱。TRIzol 处理的 SH-SY5Y 细胞产生的蛋白质 2-DE 斑点模式与等量未经处理的样品相似。随后使用肽质量指纹图谱成功鉴定了 TRIzol 处理的蛋白质。TRIzol 暴露既不改变十二烷基硫酸钠 (SDS) 凝胶中提取的肌红蛋白的质量,也不改变胶内胰蛋白酶消化产生的肌红蛋白肽的质量。这些发现表明,用 TRIzol 分离的蛋白质仍然适用于蛋白质组学分析。

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